PTPH1 cooperates with vitamin D receptor to stimulate breast cancer growth through their mutual stabilization.

Tyrosine phosphorylation is tightly regulated by protein tyrosine kinases (PTKs) and protein tyrosine phosphatases (PTPs), and has a critical role in malignant transformation and progression. Although PTKs have a well-established role in regulating breast cancer growth, contribution of PTPs remains mostly unknown. Here, we report that the tyrosine phosphatase PTPH1 stimulates breast cancer growth through regulating vitamin D receptor (VDR) expression. PTPH1 was shown to be overexpressed in 49% of primary breast cancer and levels of its protein expression positively correlate with the clinic metastasis, suggesting its oncogenic activity. Indeed, PTPH1 promotes breast cancer growth by a mechanism independent of its phosphatase activity, but dependent of its stimulatory effect on the nuclear receptor VDR protein expression and depletion of induced VDR abolishes the PTPH1 oncogenic activity. Additional analyses showed that PTPH1 binds VDR and increases its cytoplasmic accumulation, leading to their mutual stabilization and stable expression of a nuclear localization-deficient VDR abolishes the growth-inhibitory activity of the receptor independent of 1,25-dihydroxyvitamin D3. These results reveal a new paradigm in which a PTP may stimulate breast cancer growth through increasing cytoplasmic translocation of a nuclear receptor, leading to their mutual stabilization.

Plant regeneration from protoplasts isolated from embryogenic calli of the forage legume Astragalus melilotoides Pall.

An efficient and reproducible protocol is described for the regeneration of Astragalus melilotoides protoplasts isolated from hypocotyl-derived embryogenic calli. Maximum protoplast yield (11.74 +/- 0.6x10(5)/g FW) and viability (87.07 +/- 2.8%) were achieved using a mixture of 2% (w/v) Cellulase Onozuka R10, 0.5% (w/v) Cellulase Onozuka RS, 0.5% (w/v) Macerozyme R10, 0.5% (w/v) Hemicellulase, and 1% (w/v) Pectinase, all dissolved in a cell protoplast wash (CPW) salt solution with 13% (w/v) sorbitol. First divisions occurred 3-7 days following culture initiation. The highest division frequency (9.86 +/- 0.68%) and plating efficiency (1.68 +/- 0.05%) were obtained in solid-liquid medium (KM8P) supplemented with 1.0 mg/l 2,4-dichlorophenoxyacetic acid, 0.5 mg/l 6-benzylaminopurine (BA), 0.2 mg/l kinetin, 0.2 M glucose, 0.3 M mannitol and 500 mg/l casein hydrolysate. Upon transfer to MS medium with 0.5 mg/l alpha-naphthaleneacetic acid and 1-2 mg/l BA, the protoplast-derived calli produced plantlets via somatic embryogenesis (56.3 +/- 4.1%) and organogenesis (21.6 +/- 0.6%). Somatic embryos or adventitious shoots developed into well-rooted plantlets on MS medium without any plant growth regulators or supplemented with 3.0 mg/l indole-3-butyric acid, respectively. About 81% of the regenerants survived in soil, and all were normal with respect to morphology and growth characters.

Heart diseases following radiotherapy.

Three women had cardiac complaints 10-17 years after a radical mastectomy and radiotherapy for cancer of the left breast. Their ages were 42, 43 and 57 years, respectively. The first patient had a precocious coronary artery disease (CAD). She had segmental stenosis in the middle and distal portions of the left main coronary artery and pericardial fibrosis. Symptoms and signs of severe pericardial constriction became the problem after bypass surgery. She died 1 year later due to cardiorespiratory failure. The second case had sick sinus syndrome presenting a prolonged pause for 12 seconds alternating with tachy- and bradyarrhythmias, a complete atrio-ventricular block in association with pericardial effusion plus aortic and mitral valvular regurgitation. The condition improved after the implantation of a permanent pacemaker. To date, this is a very rare complication of radiation induced panconduction disturbance. The third patient had significant stenosis in the proximal portion of the left anterior descending coronary artery and myocardial fibrosis was proven by an endocardial biopsy. She had an uneventful course after medical treatment.